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Morphology and SSR fingerprinting of newly developedCynara cardunculus genotypes exploitable as ornamentals
Sergio Lanteri • Ezio Portis • Alberto Acquadro •
Rosario P. Mauro • Giovanni Mauromicale
Received: 18 May 2011 / Accepted: 5 August 2011 / Published online: 19 August 2011
� Springer Science+Business Media B.V. 2011
Abstract The species Cynara cardunculus includes
the globe artichoke (var. scolymus), the cultivated
cardoon (var. altilis) and the wild cardoon (var.
sylvestris). The three taxa are sexually compatible
and originate fertile F1 progenies, which, given the
high heterozygosity of the species, are highly segre-
gating. We report the characterization of two F1
populations, one bred from a cross between globe
artichoke and cultivated cardoon, and the other
between globe artichoke and wild cardoon. Both
populations featured a wide array of phenotypes in
relation to several traits, and some of the newly
developed genotypes are of interest for the orna-
mental market. The two populations were genotyped
at 50 microsatellite (SSR) loci: in the globe arti-
choke 9 wild cardoon and globe artichoke 9 culti-
vated cardoon progenies 116 and 97 alleles were
respectively detected. SSR pattern scores were used
to produce an UPGMA dendrogram and a PCoA plot.
A set of nine SSR loci, evenly dispersed across the
genome, was shown to be sufficient to unambigu-
ously identify each segregant. The molecular finger-
printing is useful for establishing the true to type
correspondence of propagative materials in nurseries
and ensures the effective correspondence between the
real and the declared identity of a clone.
Keywords Cynara cardunculus � Ornamentals �Phenotypic diversity � Molecular fingerprinting
Introduction
The Asteraceae family includes a number of popular
ornamentals, such as ageratum, aster, chrysanthe-
mum, dahlia, marigold, zinnia, calendula, and ger-
bera. Other members of the family have utility as leaf
vegetables (lettuce, endive), and a further group is
exploited as a source of comestible oil (sunflower,
safflower). Cynara cardunculus var. scolymus (the
globe artichoke), along with its close relatives the
cultivated (var. altilis) and the wild (var. sylvestris)
cardoon, also belongs to this botanical group.
The immature capitula of the globe artichoke are
for the most part consumed as a vegetable, and as a
result, most commercial varieties are classified on the
basis of capitulum appearance. Some varieties are
particularly suitable for fresh consumption, but others
are more appropriate for industrial transformation
(Lanteri et al. 2004a; Lanteri and Portis 2008; Mauro
S. Lanteri � E. Portis (&) � A. Acquadro
Di.Va.P.R.A. Plant Genetics and Breeding, University of
Torino, via L. da Vinci 44, 10095 Grugliasco (Torino),
Italy
e-mail: ezio.portis@unito.it
R. P. Mauro � G. Mauromicale
Dipartimento di Scienze Agronomiche, Agrochimiche e
delle Produzioni Animali—sez. Scienze Agronomiche,
University of Catania, via Valdisavoia 5, 95123 Cantania,
Italy
123
Euphytica (2012) 184:311–321
DOI 10.1007/s10681-011-0509-8
et al. 2011). The cultivated cardoon is mainly grown
in Southern Europe, where its young leaves form an
ingredient of certain traditional dishes. DNA finger-
printing has confirmed that the two cultivated forms
must have evolved independently through anthropo-
genic selection from the wild cardoon (Lanteri et al.
2004b; Portis et al. 2005a); and the assumption is that
the most likely location of the earliest selection
activity was in Sicily (Mauro et al. 2009).
Other than as a vegetable, the species has a wide
range of uses, its lignocellulosic biomass has been
proposed as a potential source of bioenergy, its seed
oil is similar in composition to that of both safflower
and sunflower (Foti et al. 1999; Ierna and Mauromi-
cale 2010; Maccarone et al. 1999), and the plant
produces a number of pharmaceutically active com-
pounds, like phenolic esters, sequiterpenes, and inulin
(Comino et al. 2007, 2009; Lattanzio et al. 2009;
Lombardo et al. 2010; Menin et al. 2010; Pandino
et al. 2010).
The various C. cardunculus taxa feature a range of
plant architecture. The foliage varies in colour from
green to ash–grey, plant height can reach 3 m, there
is a variable amount of branching (Porceddu et al.
1976), and the number of inflorescences (capitula)
per plant ranges from 5 to 15 in the globe artichoke
and from 30 to 40 in cultivated cardoon. The main
capitulum is invariably the largest. The immature
capitula are polymorphic with respect to size and
shape; some develop spiny outer bracts of various
shades of green, and which later during development
may turn purple (Cravero et al. 2005; Pochard et al.
1969). The mature capitulum can be white or violet.
Beyond all the possible cited uses, C. cardunculus
is also exploited as ornamental, both as garden plants
and as cut flowers (Cocker 1967): fresh specimens
have a long vase life, while dried forms are popular in
floral arrangements. The use of the species as
ornamental is increasing since, notwithstanding the
assortment of ornamental crops is already very large,
novelty are constantly in demand by consumers.
The highly heterozygotic nature of the species
produces a wide range of phenotypes, especially
among progeny of crosses between the different taxa
(Cravero et al. 2005; Foury 1969; Lopez-Anido et al.
1998; Mauromicale and Ierna 2000).
Molecular markers represent a reliable alternative to
morphological descriptors for varietal identification,
since they are not influenced by the growing
environment, by the developmental stage of the plant,
or by the identity of the tissue (Collard et al. 2005).
Microsatellites (SSRs) are particularly suited to DNA
fingerprinting as well as for identifying potential
parental genotypes for highly segregating populations
development, given their robustness and informative-
ness. Over the last few years, a substantial set of globe
artichoke SSR assays has been developed (Acquadro
et al. 2003, 2005a, b, 2009), and these have been used to
assess genetic diversity (Mauro et al. 2009; Portis et al.
2005a, b, c) and to construct genetic linkage maps
(Lanteri et al. 2006; Portis et al. 2009).
Here we report on phenotypic diversity released in
progenies obtained by crossing a genotype of globe
artichoke with both one of cultivated and one of wild
cardoon, as well as on the SSR-based molecular
fingerprinting of the 188 newly generated genotypes,
with a particular focus on segregants showing
potential for ornamental use.
Materials and methods
Plant material and DNA isolation
Two F1 populations were created, both involving the
globe artichoke clone ‘Romanesco C3’ as female; the
male parents were the cultivated cardoon genotype
‘Altilis 41’ (Progeny 1), and the wild cardoon
accession ‘Creta 4’ (Progeny 2). ‘Romanesco C3’ is
a late-maturing variety, which forms large purple–
green capitula (each weighing up to 400 g); its
mature capitulum develops violet coloured florets.
‘Altilis 41’ is a selection made by the University of
Catania on the basis of its high biomass yield
potential; its foliage is grey and its florets white.
‘Creta 4’ was collected from a population in Crete; it
produces a large number of capitula, forms green–
violet bracts and violet florets. A few days before
anthesis, the outer bracts of the parental capitula were
removed and the remaining structure bagged. Pollen
was collected from the male parent and stored at
3 ± 1�C for up to 3 days. Prior to the hybridization
itself, the female capitulum was rinsed in tap water,
and after 2 h the pollen was applied with a soft brush.
The bag was then replaced over the capitulum and
kept in place until the achenes had matured
(*40 days). Ripe achenes were extracted from dry
heads with a mini-thresher.
312 Euphytica (2012) 184:311–321
123
The mature achenes were germinated in moist
peat, and after 50 days, surviving seedlings (154 from
each cross), which by then had developed four true
leaves, were transplanted into the field, each spaced
1 m from its nearest neighbour. DNA was extracted
from the young plants following (Lanteri et al. 2001),
and used to confirm hybridity by SSR genotyping. A
selection of 188 true hybrids (94 from Progeny 1 and
94 from Progeny 2) was made from the confirmed
ones.
Assessment of morphology and segregations
of traits
The morphology of the hybrid individuals and three
vegetatively propagated plants of each parental
genotype was assessed over two seasons. The
assessment included traits of interest for ornamental
use like the measurement of plant height, the distance
between the first basal leaf and the upper tip of the
main capitulum, the number of days to flowering, the
duration of flowering (the number of days between
the blooming of the main and the last-produced
capitulum), capitulum colour, floret colour, the
number of capitula per plant, the maximum longitu-
dinal, and transverse diameter of the fully developed
main capitulum, and a main capitulum shape index
(the ratio between the two previously mentioned
diameters).
The goodness-of fit between observed and
expected segregation data according to previously
proposed inheritance models for the heads colour,
florets colour, and presence of spines was assessed
using the chi-square (v2) test.
SSR fingerprinting
An initial set of 93 genomic SSR assays developed in
our laboratory (Acquadro et al. 2003, 2005a, b, 2009)
was used to establish informativeness by amplifying
the DNA of the three parents and six plants of each
progeny. Each 20 ll PCR contained 10 ng genomic
DNA, 10 mM Tris–HCl (pH 8.3), 50 mM KCl,
2.5 mM MgCl2, 0.5 U Taq polymerase, 0.2 mM
dNTP, 200 nM unlabelled reverse primer and
200 nM IRD700-labelled forward primer. The touch-
down PCR protocol reported by Acquadro et al.
(2005a) was applied. The resulting amplicons were
separated by denaturing 6% polyacrylamide gel
electrophoresis using a LI-COR Gene ReadIR 4200
device, as described by Jackson and Matthews
(2000).
Fifty primer pairs identifying polymorphism in
both segregating populations were applied to the full
set of genotypes in study, following the protocol
reported above.
Amplicon sizes were estimated from the migration
of an IRD700-labelled 50–350 bp ladder. The SSR
data were collected by e-Seq software (DNA
Sequencing and Analysis Software) v3.0 and ana-
lysed using the GenAlex Excel package (Peakall and
Smouse 2006). A co-phenetic distance matrix was
generated as described by Smouse and Peakall (1999)
and used to construct a UPGMA-based dendrogram
(Sneath and Sokal 1973) within the NTSYS software
package v2.10 (Rohlf 1998). A principal coordinate
analysis (PCoA) was then performed, based on the
triangular matrix of genetic similarity estimates, and
the two axes were plotted graphically, according to
the extracted eigenvectors.
The minimum number of SSR loci needed to fully
discriminate all individuals was determined first by
selecting the set of most informative SSRs; subse-
quently, a second set was identified on the basis of the
location of the SSRs in different linkage groups on
the reference genetic linkage map (Portis et al. 2009).
Mantel tests (Mantel 1967) were performed to
establish correlations between the similarity matrices
generated by the two sets of SSRs with the one
generated from the complete data set.
Results and discussion
Phenotypic variation within the F1 populations
The two progeny sets showed an astonishing pheno-
typic variation (Fig. 1), with some individuals, the
result of specific events of chromosomal segregation
and recombination, displaying aspects of morphology
not previously observed in either cultivated or wild
types; some of these may have high potential in the
context of developing ornamental varieties of the
species.
Progeny 1 showed the most variability in terms of
flowering time (203–242 days), vegetative growth,
plant height (56–127 cm) and the number of capitula
per plant (1–32) (Table 1). Plant height may be a trait
Euphytica (2012) 184:311–321 313
123
Fig. 1 Examples of the phenotypic variation released in two inter-subspecies hybrid populations. White flower of the cultivated
cardoon parent are also included
314 Euphytica (2012) 184:311–321
123
of interest for garden plants but not for cut flowers;
on the other hand both branching of the floral stem
and the duration of flowering period, which varied
from 48 to 55 days (Table 1) in both progenies, are
key traits for both end-uses, since they both influence
the profitability of a genotype grown for cuttings as
well as its aesthetic value in a garden.
In both sets of progenies, capitulum shape was
either round (shape index \1.1) or partially elongated
and conical (shape index 1.2–2.0) (Table 1; Fig. 1).
Both the longitudinal and transverse diameter of the
capitulum were variable (in Progeny 1, the former by
6.0 cm and the latter by 4.6 cm; in Progeny 2 by,
respectively 4.2 and 4.8 cm; see Table 1). Consider-
able variation was also released for the intensity of
capitulum pigmentation, a character known to be
particularly sensitive to temperature within cultivated
germplasm.
The current genetic model for capitulum colour
assumes the presence of two dominant genes, P and
U (Cravero et al. 2005): P_ allows anthocyanin
production, resulting in purple bracts, while pp
inhibits anthocyanin production resulting in green
bracts; U_ results in an uneven distribution of
anthocyanin pigments encoded by P, while uu results
in an even distribution of pigment in the presence of
P. Among the two F1 populations it was possible to
identify the predicted three classes of capitulum
colouration (uneven purple, even purple, and green)
segregating overall in the expected digenic ratio of
9:4:3 (Table 2). Superimposed on this pattern was a
gradation in capitulum colour, along with streaks of
different colour intensity, suggesting the existence of
pigmentation modifier genes.
At flowering the capitula produce 600–1500
hermaphroditic florets. Anthesis begins at the periph-
ery and moves, over the subsequent 3–5 days, into
the centre. As the floret opens, the stigma elongates
through the tube of dehiscing anthers.
The blue–violet colour of the style and stigma was
monomorphic in both populations, despite the fact
that the cultivated cardoon parent’s florets are white
(Fig. 1). Floret colour is known to be a monogenic
trait in which the blue–violet type is the product of
the dominant allele at B (Basnitzki and Zohary 1994;
Foury and Aubert 1977). Hence, also on the basis of
our previous observation, the likely allelic state of the
three parental lines was BB in ‘Romanesco C3’, bb in
‘Altilis 41’ and BB or Bb in ‘Creta 4’.
Spiny and non-spiny types were represented in both
progeny sets; this trait is also under monogenic
control, with the dominant allele (Sp) specifying the
non-spiny trait (Lanteri et al. 2006). Both the globe
artichoke and cultivated cardoon parent were non-
spiny, while the wild cardoon was spiny. The 1:1 (50
spiny, 44 non-spiny) segregation among Progeny 2,
and the lack of any segregation in Progeny 1 showed
that the allelic constitution of the globe artichoke must
have been Spsp, that of the cultivated cardoon SpSp
and that of the wild cardoon spsp (Table 2). In both
progeny sets, some of the non-spiny types developed
small thorns on both the leaves and capitula, while in
others, small thorns developed only on the leaves,
leaving the apex of the bracts flat or even indented
with a small thorn in its centre (data not shown). The
absence of thorns (or, if they are present, then they are
small and soft) is clearly a positive trait in terms of
handling the plant for cut flower purposes.
Table 1 Phenotypic variation observed for seven traits in the two inter-subspecies hybrid populations [globe artichoke 9 cultivated
cardoon (Progeny 1) and globe artichoke 9 wild cardoon (Progeny 2)]
Trait Progeny 1 Progeny 2
Mean Minimum Maximum Mean Minimum Maximum
Plant height (cm) 95 56 127 54 28 89
Days to flowering 216 203 242 223 214 234
Flowering period (days) 25 7 55 22 2 48
Number of heads plant-1 7.4 1 32 5.7 1 20
Head longitudinal diameter (cm) 6.9 4.4 10.4 6.4 4.0 8.2
Head transversal diameter (cm) 5.1 3.2 7.8 4.4 2.8 7.6
Shape index 1.37 1.04 1.94 1.45 1.07 1.97
Euphytica (2012) 184:311–321 315
123
Leaf shape and upper and lower leaf surface
tomentosity were also highly variable in both progeny
sets. The former varied from deeply divided to lobed,
while the intensity of tomentosity strongly influenced
leaf colour, which ranged from an ash–grey (typical
of the wild cardoon) to deep green (like that of the
leaves of the globe artichoke) (data not shown).
SSR-based genotyping
Cross-pollination in the globe artichoke is largely
assured by protandry, but a degree of self-pollination
does occur, because the stigma remains receptive to
pollen for 4–5 days after pollen shed, and so periph-
eral florets can be fertilized by pollen from central
florets. Self-pollination can also occur via pollen
transfer between asynchronous capitula produced by
the same plant. Thus it was important to verify the
hybridity of the progeny obtained by cross-pollina-
tion, and this was achieved using SSR profiling at two
informative loci.
All 93 SSR assays initially applied generated a
profile consisting of either one or two alleles per
template, as expected from a diploid species, and the
allelic constitution. Among them, 50 loci segregated
in both F1 populations for at least one parent and
were then applied to the full set of hybrid individuals
(Table 3). Within Progeny 1, the segregation of 17
loci was consistent with a 1:1:1:1 ratio; of these, 12
displayed a four allele segregation (segregation
type \ab 9 cd[), and the other five a three allele
segregation (\ab 9 ac[). One locus was consistent
with a 1:2:1 ratio (segregation type \ab 9 ab[)
while the remaining 32 loci segregated within only
one of the parents, producing a segregation pattern
consistent with a 1:1 ratio (Table 3). Within Progeny
2, 25 loci proved to be heterozygous in both the
parents, all of them segregating in a 1:1:1:1 ratio (16
with four alleles and nine with three alleles); the other
25 loci segregated within only one of the parents (1:1
ratio). As also noted by Portis et al. (2009), the level
of heterozygosity in the cultivated cardoon was less
than in the wild cardoon, and so more of the loci
segregated only for the globe artichoke alleles
(Table 3).
The segregation analysis also revealed a number of
null alleles, which occur at SSR loci when one (or
both) primer annealing sites are lost by mutation or
Table 2 Observed and expected segregation for the heads colour, florets colour, and presence of spines in the two inter-subspecies
hybrid populations [globe artichoke 9 cultivated cardoon (Progeny 1) and globe artichoke 9 wild cardoon (Progeny 2)]
Number of individuals Globe artichoke
‘Romanesco C3’
Cultivated cardoon
‘Altilis 41’
Wild cardoon
‘Creta 4’
Progeny 1 Progeny 2
Purple-green heads X X X 53 52
Green heads 24 27
Purple heads 17 15
Expected ratio 9:4:3 9:4:3
v2 0.03 ns 0.39 ns
Putative genotype PpUu PpUu PpUu [P_U_ (9)]:[ppU_ ? ppuu (4)]:[P_uu (3)]
Violet florets X X 94 94
White florets X 0 0
Expected ratio 1:0 1:0
v2 ns ns
Putative genotype BB bb BB (or Bb) Bb BB (or BB ? Bb)
Spiny heads X 0 44
Not spiny heads X X 94 50
Expected ratio 1:0 1:1
v2 ns 0.38 ns
Putative genotype Spsp SpSp spsp SpSp ? Spsp [Spsp (1)]:[spsp (1)]
316 Euphytica (2012) 184:311–321
123
deletion (Jones et al. 1998; Holm et al. 2001;
Pemberton et al. 1995). The effect of null alleles is
to over-estimate the frequency of homozygosity,
since it becomes no longer possible to discriminate
homozygotes from heterozygotes (Pekkinen et al.
2005). In this situation, the options are either to
disregard the affected loci, to score segregation in the
same way as for a dominant marker (Rodzen and
May 2002), to attempt to redesign the primers (Shaw
et al. 1999; Van Oosterhout et al. 2004), or to adjust
allele frequencies on the basis of a global estimate of
the frequency of null alleles. However, for the present
populations, it was generally possible to identify the
presence of null alleles. Within Progeny 2, null
alleles were inferred at 12 of the 50 SSR loci. As
described in Fig. 2, seven of these loci displayed the
pattern named (a) and five pattern (b). Patterns
(c) and (d) detected monomorphic null alleles, and
thus were not taken into consideration. The null
alleles at two of the SSR loci displayed a segregation
pattern which was ambiguous (Fig. 3) and so were
excluded from any further analysis. Within Progeny
1, null alleles were identified at just five of the 50
loci, presumably as a consequence of the lower
phylogenetic distance between the two parental
genotypes. Indeed the highest similarity value
detected between pairs of genotypes in the Progeny
1 was 0.73 while in the Progeny 2 it was 0.82.
Table 3 Segregation patterns and number of polymorphic/informative alleles detected in the in two inter-subspecies hybrid pop-
ulations [globe artichoke 9 cultivated cardoon (Progeny 1) and globe artichoke 9 wild cardoon (Progeny 2)]
Segregation type Segregation
ratio
No. informative
alleles/locus
Progeny 1 Progeny 2
No.
loci
Total no.
of alleles
No.
loci
Total no.
of alleles
\ab 9 cd[ 1:1:1:1 4 12 48 16 64
\ab 9 ac[ 1:1:1:1 3 5 15 9 27
\ab 9 ab[ 1:2:1 2 1 2 0 0
\ab 9 aa[\ab 9 cc[\aa 9 ab[\aa 9 bc[
1:1 1 32 32 25 25
Total 50 97 50 116
First parent F1 segregating progeny
(segregation ratio 1:1:1:1)
Secondparent
(a)
x
(segregation ratio 1:1:1:1)(b)
x
(segregation ratio 1:1)(c)x
(segregation ratio 1:1)(d)
(ab) (cd) (ac) (ad) (bc) (bd)
(ab) (bc) (ab) (ac) (bb) (bc)
(ab) (bb) (ab) (bb)
(ab) (cc) (ac) (bc)
x
Fi d
Fig. 2 Segregation patterns which allow for the identification
of null alleles (indicated by a white band). A null allele was
inferred to be segregating when a single product or no product
was observed
First parent F1 segregating progeny
Secondparent
(segregation ratio 1:2:1)(a)
x
(segregation ratio 3:1)(b)
x
(ab) (bc) (ab) (ac)(bb) (bc)
(ab) (ab) (aa) (bb)(ab) (ab)
Fig. 3 Segregation pattern which does not allow for the
identification of null alleles (indicated by a white band). Both
homozygous and heterozygous individuals amplify only a
single product
Euphytica (2012) 184:311–321 317
123
Ro
man
esco
C3
x A
ltili
s 41
Ro
man
esco
C3
x C
reta
4
Altilis 41
Romanesco C3
Creta 4
0.25 0.40 0.55 0.70 0.85 1.00
Jaccard’s SimilarityCoefficient
Fig. 4 UPGMA-based
cluster analysis of the
segregants arising in two
inter-subspecies hybrid
populations, based on their
allelic constitution at nine
SSR loci. In the dendrogram
parental genotypes are
indicated
318 Euphytica (2012) 184:311–321
123
On the basis of the whole 50 SSR primers set it
was possible to obtain a specific molecular finger-
printing for each individual in both segregating
progenies. The initial attempt to identify the mini-
mum number of SSR loci needed to fully discrimi-
nate between all individuals was based on a selection
of 16 SSRs at which four alleles were detected in
Progeny 2 (Table 3). These were used to create a
similarity matrix between each individual; the
most similar pair was 78% similar to one another;
the correlation between this matrix and the total
similarity matrix indicated a good fit between these
two representations of the genetic relationships
(r = 0.87).
Nine of the 16 SSRs (CELMS-01, -15, -23, -24,
-37, -41, -58, and -60, and CMAL-21) were then
chosen on the basis that these loci are dispersed
across different linkage groups (Portis et al. 2009).
The similarity matrix obtained from this set was well
correlated with the one obtained using the whole data
set (r = 0.78). The most similar pair of individuals
from Progeny 1 were 90% similar to one another,
while those from Progeny 2 were 85% similar to one
another. The nine SSR loci were able to fully
discriminate all members of both progeny sets. The
UPGMA-based clustering (Fig. 4) separated the
genotypes into two major clades, corresponding to
the two F1 populations, with the common parent
(‘Romanesco C3’) at an intermediate position.
The first two axes of the PCoA scatter plot (Fig. 5)
explained, respectively, 39 and 23% of the overall
genetic variation; the former largely discriminated
between the two populations and highlighted the
higher variability present within Progeny 2. Indeed in
Progeny 2 a molecular fingerprinting of each geno-
type might be obtain by applying only seven (i.e.,
CELMS-01, -15, -24, -37, -41, -58, and CMAL-21) of
the nine selected SSRs. A further reduction in the
number of SSR loci needed for molecular finger-
printing could be possible if the focus is restricted to
types suitable for the development of ornamental
varieties.
Conclusions
Here we have reported the release of a large amount
of phenotypic diversity by crossing the globe arti-
choke with either the wild or the cultivated cardoon.
On the basis of our previous studies, due to the
heterozygotic nature of the species, highly segregat-
ing progenies may be also obtained following selfing
or by crossing spiny with non-spiny globe artichoke
varietal types; however, the amount of released
phenotypic diversity is by far less pronounced than
the one we observed in our inter-taxa progenies.
A number of the segregants displayed aspects of
morphology which have high potential as ornamen-
tals. A particular advantage of this species is that any
individual can be readily immortalised by vegetative
propagation, either by isolating basal growing shoots
or semi-dormant shoots which develop on the under-
ground stem. Indeed, a set of thirty of the most
promising genotypes has been vegetatively propa-
gated by isolating basal growing shoots, and are
currently under evaluation in different environments
with the goal of identifying the most valuable for
future marketing. Micropropagation is a further
option, as protocols for meristem culture have been
well established for the species, also starting from
meristem tips, with the goal to obtain virus free plants
for the production of sanitary controlled propagative
material (Acquadro et al. 2010; Barba et al. 2004;
Papanice et al. 2004).
A set of nine SSR loci, evenly dispersed across the
genome, was shown to be sufficient to unambiguously
Principal Component 1
-0.40 -0.15 0.10 0.35 0.60
Prin
cipa
l Com
pone
nt 2
-0.55
-0.28
0.00
0.28
0.55
Romanesco C3
Altilis 41Creta 4
Fig. 5 Principal coordinate analysis (PCoA) based on SSR-
based genotypic data, depicting the genetic relationship
between the segregants arising in two inter-subspecies hybrid
populations. Progeny 1 shown as grey circles, Progeny 2 as
white circles. The parents are indicated by triangles
Euphytica (2012) 184:311–321 319
123
identify each segregant. The DNA profiles generated
by these SSR assays will have utility in establishing the
genetic identity of vegetatively propagated materials.
References
Acquadro A, Portis E, Lanteri S (2003) Isolation of microsat-
ellite loci in artichoke (Cynara cardunculus L. var. scol-ymus). Mol Ecol Notes 3:37–39
Acquadro A, Portis E, Albertini E, Lanteri S (2005a) M-AFLP-
based protocol for microsatellite loci isolation in Cynaracardunculus L. (Asteraceae). Mol Ecol Notes 5:272–274
Acquadro A, Portis E, Lee D, Donini P, Lanteri S (2005b)
Development and characterization of microsatellite
markers in Cynara cardunculus L. Genome 48:217–225
Acquadro A, Lanteri S, Scaglione D, Arens P, Vosman B,
Portis E (2009) Genetic mapping and annotation of
genomic microsatellites isolated from globe artichoke.
Theor Appl Genet 118(8):1573–1587
Acquadro A, Papanice M, Lanteri S, Bottalico G, Portis E,
Campanale A, Finetti-Sialer M, Mascia T, Sumerano P,
Gallitelli D (2010) Production and fingerprinting of virus-
free clones in a reflowering globe artichoke. Plant Cell
Tiss Organ Cult 100(3):329–337
Anido F, Firpo I, Garcia S, Cointry E (1998) Estimation of
genetic parameters for yield traits in globe artichoke
(Cynara scolymus L.). Euphytica 103:61–66
Barba M, Di Lernia G, Babes G, Citrulli F (2004) Produzione e
conservazione di germoplasma di carciofo di tipo ro-
manesco esente da virus. Italus Hortus 11:5–10
Basnitzki J, Zohary D (1994) Breeding of seed planted arti-
choke. Plant Breed Rev 12:253–269
Cocker H (1967) Il carciofo pianta ornamentale. In: Medica M
(ed) I International Congress on Artichoke. Minerva
Medica, Bari, pp 313–317
Collard B, Jahufer M, Brouwer J, Pang E (2005) An intro-
duction to markers, quantitative trait loci (QTL) mapping
and marker-assisted selection for crop improvement: the
basic concepts. Euphytica 142:169–196
Comino C, Lanteri S, Portis E, Acquadro A, Romani A,
Hehn A, Larbat R, Bourgaud F (2007) Isolation and
functional characterization of a cDNA coding a hydro-
xycinnamoyltransferase involved in phenylpropanoid
biosynthesis in Cynara cardunculus L. BMC Plant Biol
7:14
Comino C, Hehn A, Moglia A, Menin B, Bourgaud F, Lanteri
S, Portis E (2009) The isolation and mapping of a novel
hydroxycinnamoyltransferase in the globe artichoke
chlorogenic acid pathway. BMC Plant Biol 9:30
Cravero V, Picardi L, Cointry E (2005) An approach for
understanding the heredity of two quality traits (head
color and tightness) in globe artichoke (Cynara scolymusL.). Genet Mol Biol 28:431–434
Foti S, Mauromicale G, Raccuia S, Fallico B, Fanella F,
Maccarone E (1999) Possible alternative utilization of
Cynara spp. I. Biomass, grain yield and chemical com-
position of grain. Ind Crop Prod 10:219–228
Foury C (1969) Etude de la biologie florale de l’artichaut
(Cynara scolymus L.). Application a la selection 2 partie.
Etude des descendances obtenues en fecondation con-
trolee. Ann Amelior Plantes 19:23–52
Foury C, Aubert S (1977) Observation preliminares sur la
presence et la repartition de pigments anthocyaniques
dans un mutant d’artichaut (Cynara scolymus L.) a fleurs
blanches. Ann Amelior Plantes 27(5):603–612
Holm L, Loeschcke V, Bendixen C (2001) Elucidation of the
molecular basis of a null allele in a rainbow trout
microsatellite. Mar Biotechnol 3:555–560
Ierna A, Mauromicale G (2010) Cynara cardunculus L.
genotypes as a crop for energy purposes in a Mediterra-
nean environment. Biomass Bioenerg 34(5):754–760
Jackson JA, Matthews D (2000) Modified inter-simple
sequence repeat PCR protocol for use in conjunction with
the LI-COR gene ImagIR(2) DNA analyzer. Biotechni-
ques 28:914–916
Jones A, Stockwell C, Walker D, Avise J (1998) The molecular
basis of a microsatellite null allele from the white sands
pupfish. J Hered 89:339–342
Lanteri S, Portis E (2008) Globe Artichoke and Cardoon. In:
Springer (ed) Vegetables I, vol 1. Springer, New York,
pp 49–74
Lanteri S, Di Leo I, Ledda L, Mameli M, Portis E (2001)
RAPD variation within and among populations of globe
artichoke cultivar ‘Spinoso sardo’. Plant Breeding 120:
243–246
Lanteri S, Acquadro A, Saba E, Portis E (2004a) Molecular
fingerprinting and evaluation of genetic distances among
selected clones of globe artichoke (Cynara cardunculus L.
var. scolymus L.). J Hortic Sci Biotech 79:863–870
Lanteri S, Saba E, Cadinu M, Mallica G, Baghino L, Portis E
(2004b) Amplified fragment length polymorphism for
genetic diversity assessment in globe artichoke. Theor
Appl Genet 108:1534–1544
Lanteri S, Acquadro A, Comino C, Mauro R, Mauromicale G,
Portis E (2006) A first linkage map of globe artichoke
(Cynara cardunculus var. scolymus L.) based on AFLP,
S-SAP, M-AFLP and microsatellite markers. Theor Appl
Genet 112:1532–1542
Lattanzio V, Kroon PA, Linsalata V, Cardinali A (2009) Globe
artichoke: a functional food and source of nutraceutical
ingredients. J Funct Food 1:131–144
Lombardo S, Pandino G, Mauromicale G, Knodler M, Carle R,
Schieber M (2010) Influence of genotype, harvest time
and plant part on polyphenolic composition of globe
artichoke [Cynara cardunculus var. scolymus (L.) Fiori].
Food Chem 119:1175–1181
Maccarone E, Fallico B, Fanella F, Mauromicale G, Raccuia S,
Foti S (1999) Possible alternative utilization of Cynaraspp. II. Chemical characterization of their grain oil. Ind
Crop Prod 10(1999):229–237
Mantel N (1967) The detection of disease clustering and a
generalized regression approach. Cancer Res 27:209–220
Mauro R, Portis E, Acquadro A, Lombardo S, Mauromicale G,
Lanteri S (2009) Genetic diversity of globe artichoke
landraces from Sicilian small-holdings: implications for
evolution and domestication of the species. Conserv
Genet 10:431–440
320 Euphytica (2012) 184:311–321
123
Mauro R, Lombardo S, Longo AMG, Pandino G, Mauromicale
G (2011) New cropping designs of globe artichoke for
industrial use. Ital J Agron 6:e8
Mauromicale G, Ierna A (2000) Panorama varietale e migliora-
mento genetico del carciofo. Informatore agrario 26:39–45
Menin B, Comino C, Moglia A, Dolzhenko Y, Portis E, Lanteri
S (2010) Identification and mapping of genes related to
caffeoylquinic acid synthesis in Cynara cardunculus L.
Plant Sci 178:338–347
Pandino G, Courts F, Lombardo S, Mauromicale G, William-
son G (2010) Caffeoylquinic acids and flavonoids in the
immature inflorescence of globe artichoke, wild cardoon,
and cultivated cardoon. J Agric Food Chem 58:1026–1031
Papanice MA, Campanale A, Bottalico G, Sumerano P, Gal-
litelli D (2004) Production of virus-free artichoke germ-
plasm cv Brindisino (Cynara scolymus L.; Apulia). Italus
Hortus 11(5):11–15
Peakall R, Smouse P (2006) GENALEX 6: genetic analysis in
excel. Population genetic software for teaching and
research. Mol Ecol Notes 6:288–295
Pekkinen M, Varvio S, Kulju K, Karkkainen H, Smolander S,
Vihera-Aarnio A, Koski V, Sillanpaa M (2005) Linkage
map of birch, Betula pendula Roth, based on microsatel-
lites and amplified fragment length polymorphisms.
Genome 48:619–625
Pemberton J, Slate J, Bancroft D, Barrett J (1995) Non-
amplifying alleles at microsatellite loci—a caution for
parentage and population studies. Mol Ecol 4:249–252
Pochard E, Foury C, Chambonet D (1969) Il miglioramento
genetico del carciofo. Proceedings the 1 Congresso
Internazionale sul carciofo–Bari–Italy, pp 117-155
Porceddu E, Dellacecca V, Bianco V (1976) Classificazione
numerica di cultivar di carciofo. Proceedings II Interna-
tional Congress on Artichoke, Ed Minerva Medica, Torino
pp 1105-1119
Portis E, Barchi L, Acquadro A, Macua J, Lanteri S (2005a)
Genetic diversity assessment in cultivated cardoon by
AFLP (Amplified Fragment Length Polymorphism) and
microsatellite markers. Plant Breed 124:299–304
Portis E, Acquadro A, Comino C, Mauromicale G, Saba E,
Lanteri S (2005b) Genetic structure of island populations
of wild cardoon [Cynara cardunculus L. var. sylvestris(Lamk) Fiori] detected by AFLPs and SSRs. Plant Sci
169:199–210
Portis E, Mauromicale G, Barchi L, Mauro R, Lanteri S
(2005c) Population structure and genetic variation in
autochthonous globe artichoke germplasm from Sicily
Island. Plant Sci 168:1591–1598
Portis E, Mauromicale G, Mauro R, Acquadro A, Scaglione D,
Lanteri S (2009) Construction of a reference molecular
linkage map of globe artichoke (Cynara cardunculus var.
scolymus). Theor Appl Genet 120(1):59–70
Rodzen J, May B (2002) Inheritance of microsatellite loci in
the white sturgeon (Acipenser transmontanus). Genome
45:1064–1076
Rohlf FJ (1998) NTSYSpc Version 2.0: User Guide. Applied
Biostatistics Inc
Shaw P, Turan C, Wright J, O’Connell M, Carvalho G (1999)
Microsatellite DNA analysis of population structure in
Atlantic herring (Clupea harengus), with direct compari-
son to allozyme and mtDNA RFLP analyses. Heredity
83:490–499
Smouse P, Peakall R (1999) Spatial autocorrelation analysis of
individual multiallele and multilocus genetic structure.
Heredity 82:561–573
Sneath PHA, Sokal RR (1973) Numerical taxonomy—the
principles and practice of numerical classification. W, H.
Freeman, San Francisco
Van Oosterhout C, Hutchinson W, Wills D, Shipley P (2004)
MICRO-CHECKER: software for identifying and cor-
recting genotyping errors in microsatellite data. Molecular
Ecology Notes, vol 4. Wiley, New York, pp 535–538
Euphytica (2012) 184:311–321 321
123